Biological sample quantitative detection method based on optical disc

Abstract

The invention discloses a biological sample quantitative detection method based on an optical disc, and belongs to the technical field of biosensors, and the method comprises the following steps: reacting a solution to be detected of a mixed system with an insoluble precipitate based on the inhibition or promotion effect of the solution to be detected on the insoluble precipitate solution, and preparing a suspension for the residual insoluble precipitate in the reaction; the method comprises the following steps: after activating the surface of an optical disc full of data, dropwise adding an insoluble precipitate suspension corresponding to a mixed system solution to be detected or a single system solution to be detected on a specified radius of the surface of the optical disc to form sample dropping points; after moisture in the sample dropping point is completely evaporated, reading data in the optical disc, obtaining data reading errors, and calculating a corresponding LDC count; and substituting the LDC count into a pre-calibrated standard curve or relational expression to obtain the concentration of the solution to be detected. According to the invention, the efficiency and universality of quantitative detection of substances in the solution can be improved, and the detection cost and the detection complexity are reduced.

Description

Technical Field

[0001] The present invention belongs to the technical field of biosensors, and more specifically, relates to a biological sample quantitative detection method based on an optical disk. Background Art

[0002] At present, biochemical analysis has become an important task in the field of analytical science. Existing methods such as mass spectrometry, spectral analysis, chromatography and enzyme-linked immunosorbent assay (ELISA) have made significant progress in accuracy, sensitivity, detection throughput and automation. However, traditional biochemical analysis technology is limited by expensive instruments and professional operations, and is mainly confined to laboratories in developed areas. Through the "adaptive use" strategy, that is, using the physical mechanism of everyday objects, the cost of detection can be reduced, the operation can be simplified, and the accessibility and versatility of detection can be improved, which is of great significance for early on-site biomedical analysis, especially in resource-limited areas.

[0003] In recent years, consumer electronic products such as optical drives have been adapted as tools for on-site analysis and point-of-care testing (POCT), especially in the field of biochemical testing. The precision optics and servo systems of optical drives enable optical discs to serve as substrates for biochemical reactions, thus developing a variety of biochemical detection strategies based on optical disc technology. Although these strategies are innovative, they usually require customization or modification of optical drives, optical discs, or software, which to some extent limits their widespread application. In addition, problems such as complex quantitative dispensing operations, high usage barriers, insufficient integration, large reagent consumption, slow detection speed, and high cost remain challenges facing current technologies.

[0004] To address the above issues, researchers have proposed a strategy that does not require hardware modification or new software development. However, the process of constructing a reaction system on the surface of a Blu-ray Disc (BD) is still cumbersome and time-consuming, requiring multiple steps of reaction (such as surface activation, probe fixation, sample addition and blocking) and additional labeling reagents (such as silver staining). This not only increases operational errors, but also raises the requirements for the professionalism of operators, thereby limiting the promotion and application of these technologies to a certain extent. Summary of the invention

[0005] In view of the defects and improvement needs of the prior art, the present invention provides a method for quantitative detection of biological samples based on optical disks, which aims to improve the efficiency and versatility of quantitative detection of substances in solutions and reduce detection costs and detection complexity.

[0006] To achieve the above object, according to one aspect of the present invention, a biological sample quantitative detection method based on an optical disc is provided, which is used to detect the concentration of a solution to be tested; the solution to be tested is a soluble target solution or an insoluble precipitate suspension, and the soluble target of the soluble target solution and the insoluble precipitate in the insoluble precipitate suspension will not damage the surface of the optical disc; the optical disc is full of data; the quantitative detection method comprises:

[0007] After the optical disc is activated, the solution to be tested is dripped on the surface of the optical disc at a specified radius to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state;

[0008] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0009] Determine the concentration corresponding to the calculated LDC counts according to a pre-calibrated standard curve to obtain the concentration of the solution to be tested;

[0010] Among them, when the test solution is a soluble target solution, the standard curve is obtained by fitting the LDC counts corresponding to soluble target solutions of different concentrations; when the test solution is an insoluble precipitate suspension, the standard curve is obtained by fitting the LDC counts corresponding to insoluble precipitate suspensions of different concentrations.

[0011] Furthermore, the calibration method of the standard curve includes:

[0012] Prepare test solutions with different concentrations as standard solutions;

[0013] After the optical disc is activated, the standard solution is evenly dripped on the surface of the optical disc along a specified radius to obtain a sample drop point corresponding to each standard solution;

[0014] After the water in all sample drop points is completely evaporated, the data in the CD is read to obtain the data reading error corresponding to each sample drop point, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution;

[0015] Fitting was performed based on the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the corresponding relationship between the concentration and the LDC count.

[0016] According to another aspect of the present invention, a method for quantitative detection of biological samples based on an optical disc is provided, which is used to detect the concentration of a solution to be tested, wherein the solution to be tested is a soluble target solution; the optical disc is filled with data; the quantitative detection method comprises:

[0017] Step S1: mixing the test solution with an excess of insoluble precipitate suspension, so that the soluble target in the test solution and the insoluble precipitate in the insoluble precipitate suspension fully react; the soluble target can react with the insoluble precipitate and produce a soluble product after the reaction; the insoluble precipitate will not damage the surface of the optical disc;

[0018] Step S2: centrifuging the mixed solution;

[0019] Step S3: after removing the supernatant from the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new insoluble precipitate suspension;

[0020] Step S4: After the optical disc is activated, the insoluble precipitate suspension obtained in step S3 is dripped onto the surface of the optical disc at a specified radius to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state;

[0021] Step S5: after the water in the sample droplet point is completely evaporated, the data in the optical disc is read to obtain the data reading error therein, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0022] Step S6: Substitute the LDC count obtained in step S5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

[0023] Furthermore, the calibration method of the relationship between the LDC count and the concentration of the solution to be tested includes:

[0024] Prepare test solutions with different concentrations as standard solutions;

[0025] Each standard solution is mixed with an insoluble precipitate suspension of equal concentration and excess, so that the soluble target in each standard solution and the insoluble precipitate in the insoluble precipitate suspension can fully react;

[0026] Each mixed solution is centrifuged separately;

[0027] After removing the supernatant from the mixed solution after centrifugation, deionized water was added to the remaining solution to obtain a new insoluble precipitate suspension corresponding to each standard solution;

[0028] After the optical disc is activated, the insoluble precipitate suspension corresponding to each standard solution is evenly dropped on the surface of the optical disc along a specified radius to obtain a sample drop point corresponding to each standard solution;

[0029] After the water in all sample drop points is completely evaporated, the data in the CD is read to obtain the data reading error corresponding to each sample drop point, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution;

[0030] The concentration of each standard solution and the corresponding LDC count were fitted to obtain the relationship between the LDC count and the concentration of the solution to be tested.

[0031] According to another aspect of the present invention, a method for quantitative detection of biological samples based on an optical disc is provided, which is used to detect the concentration of a solution to be tested, wherein the solution to be tested is a soluble target solution; the optical disc is filled with data; the quantitative detection method comprises:

[0032] Step T1: mixing the test solution, the insoluble precipitate suspension and the third reactant solution, so that the insoluble precipitate in the insoluble precipitate suspension fully reacts with the third reactant in the third reactant solution; the insoluble precipitate can react with the third reactant to produce a soluble product after the reaction, and the insoluble precipitate suspension is in excess relative to the third reactant solution; the test solution will inhibit the reaction of the third reactant with the insoluble precipitate, and the higher the concentration of the test solution, the stronger the inhibitory effect; the insoluble precipitate will not damage the surface of the optical disc;

[0033] Step T2: centrifuging the mixed solution;

[0034] Step T3: after removing the supernatant from the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new insoluble precipitate suspension;

[0035] Step T4: After the optical disc is activated, the insoluble precipitate suspension obtained in step T3 is dripped onto the surface of the optical disc at a specified radius to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state;

[0036] Step T5: After the water in the sample dripping clock is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0037] Step T6: Substitute the LDC count obtained in step T5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

[0038] Furthermore, the calibration method of the relationship between the LDC count and the concentration of the solution to be tested includes:

[0039] Prepare test solutions with different concentrations as standard solutions;

[0040] For each standard solution, mix it with the insoluble precipitate suspension and the third reactant solution, so that the insoluble precipitate and the third reactant fully react; the concentration of the insoluble precipitate suspension mixed with each standard solution is equal, and the concentration of the third reactant solution mixed with each standard solution is equal;

[0041] Each mixed solution is centrifuged separately;

[0042] After removing the supernatant from the mixed solution after centrifugation, deionized water was added to the remaining solution to obtain a new insoluble precipitate suspension corresponding to each standard solution;

[0043] After the optical disc is activated, the insoluble precipitate suspension corresponding to each standard solution is evenly dropped on the surface of the optical disc along a specified radius to obtain a sample drop point corresponding to each standard solution;

[0044] After the water in all sample drop points is completely evaporated, the data in the CD is read to obtain the data reading error corresponding to each sample drop point, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution;

[0045] The concentration of each standard solution and the corresponding LDC count were fitted to obtain the relationship between the LDC count and the concentration of the solution to be tested.

[0046] According to another aspect of the present invention, a method for quantitative detection of biological samples based on an optical disc is provided, which is used to detect the concentration of a bacterial solution; the optical disc is filled with data; the quantitative detection method comprises:

[0047] After the optical disc is activated, a bacterial solution is dripped on the surface of the optical disc at a specified radius to form a sample drop point; the activation treatment changes the surface of the optical disc from a hydrophobic state to a hydrophilic state; after the activation treatment, the ability of the optical disc surface to adsorb bacteria will be enhanced;

[0048] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0049] Determine the concentration corresponding to the calculated LDC count according to a pre-calibrated standard curve to obtain the concentration of the bacterial solution;

[0050] The standard curve is obtained by fitting the LDC counts corresponding to bacterial solutions of different concentrations.

[0051] Furthermore, the calibration method of the standard curve includes:

[0052] Prepare bacterial solutions with different concentrations as standard solutions;

[0053] After the optical disc is activated, the standard solution is evenly dropped on the surface of the optical disc along a specified radius to obtain a sample drop point corresponding to each standard solution;

[0054] After the water in all sample drop points is completely evaporated, the data in the CD is read to obtain the data reading error corresponding to each sample drop point, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution;

[0055] Fitting was performed based on the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the corresponding relationship between the concentration and the LDC count.

[0056] Furthermore, in the above optical disc-based biological sample quantitative detection method, the activation process comprises:

[0057] The optical disc is immersed in a NaOH solution for a preset time to generate reactive carboxyl groups on its surface, thereby changing from a hydrophobic state to a hydrophilic state.

[0058] In general, through the above technical solutions conceived by the present invention, a new "direct spot sampling-drying" (drip-dry strategy, 2D) strategy is proposed, that is, the sample solution is directly dripped on the surface of the activated optical disc to form a sample drop point, and after the water in the sample drop point is completely evaporated, the data is read, and the quantitative detection result is obtained based on the data reading error. The relevant detection principle is: the substance in the sample drop point is deposited on the surface of the optical disc to form a covering area after the water is completely evaporated, thereby causing the laser to be unable to reflect and causing a data reading error after the optical drive reads the data, and the data reading error has a calibrable quantitative relationship with the sample concentration; the data reading error caused by the sample drop point can be obtained using the existing optical disc quality diagnosis software, and then the quantitative detection can be completed.

[0059] Based on the proposed "direct spotting-drying" strategy, the present invention adopts the "direct spotting-drying" strategy for solutions that will not damage the surface of the optical disc. There is no need to customize or modify the optical drive, optical disc or software, and there is no need for complex operations such as probe fixation and sealing. There is no need to use additional labeling reagents such as silver staining. The operation is simple and convenient, the detection efficiency is high, the detection complexity is low and the cost is low. At the same time, there are no special operating requirements for related operations, and its versatility is greatly improved.

[0060] For complex mixed system solutions or solutions that may damage the surface of the optical disc, an insoluble precipitate suspension that will not damage the surface of the optical disc is introduced, and the reaction process of the insoluble precipitate is regulated by the soluble target in the test solution, that is, the soluble target is used to react with the insoluble precipitate, or the soluble target is used to inhibit the reaction between the insoluble precipitate and other reactants to regulate the dissolution process of the insoluble precipitate, so as to achieve effective purification of the insoluble precipitate, and then the concentration of the purified insoluble precipitate suspension is detected through the "direct spotting-drying" strategy, so as to indirectly achieve quantitative detection of the test solution. The entire purification process is carried out outside the optical disc, and the surface of the optical disc only needs to implement the "direct spotting-drying" strategy. Similarly, the operation is simple and convenient, with high detection efficiency, low detection complexity, low cost and high versatility.

[0061] Compared with the prior art, the present invention can achieve the following beneficial effects:

[0062] (1) When the present invention performs quantitative detection of biological samples, only a CD, a standard CD-ROM drive device and CD-ROM quality diagnosis software are needed. The relevant software and hardware are easy to obtain, which greatly reduces the detection cost (single test cost < 1 US dollar).

[0063] (2) The entire detection process, including sample processing, error reading and result analysis, can be completed within 30 minutes. The sample processing operation is simple and easy to use, providing an alternative method for rapid material analysis for early, on-site and home use.

[0064] (3) The present invention is universal and applicable to the quantification of almost all substances in a single system; in the case of a mixed system, the method is also applicable to most reactions that can produce or dissolve precipitates, converting the quantification of the target substance into the quantification of a separable and purifiable precipitate. The method can provide new ideas and methods for material analysis and research in many fields such as biomedicine, food hygiene, and environmental testing.

[0065] (4) The diameter of the laser spot on the surface of the optical disc can reach the micron level. For example, a Blu-ray disc uses a laser with a wavelength of 405 nm, and the data layer is only 0.1 mm away from the surface. The diameter of the laser spot on the surface is 138 μm. Therefore, the quantitative detection method of biological samples provided by the present invention can, in theory, have a resolution of up to the micron level. High-throughput detection can be achieved by constructing more densely arranged detection sites on the surface of the optical disc.

[0066] In general, the present invention combines daily optical drive equipment and free software to build a low-cost, efficient biological sample quantitative detection system. The system can complete the test within 30 minutes, is suitable for quantitative analysis of substances in multiple fields, and has high-throughput detection capabilities, providing a new approach for future research and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 A schematic diagram of software and hardware related to the biological sample quantitative detection method provided by the present invention;

[0068] Figure 2 The structure of the Blu-ray disc provided by the present invention and the basic principle of BD error reading are shown in FIG. 1 , a hard coating layer, 2, a cover layer, 3, an adhesive layer, 4, a recording layer, 5, a reflective layer, 6, a substrate, and 7, a protective layer.

[0069] Figure 3 A schematic diagram of the process of the optical disc quality diagnosis software used in the present invention reading and analyzing BD errors and then calculating the error rate;

[0070] Figure 4 Schematic diagram of the operation process of the "direct spotting-drying" strategy (2D strategy) in the present invention;

[0071] Figure 5 Schematic diagram of the principle and operation flow of the "direct spotting-drying" strategy provided by the present invention in three typical biological monitoring methods;

[0072] Figure 6 A schematic diagram of the data results obtained by quantitative detection of bovine serum albumin provided in Example 1 of the present invention; wherein (a) is the original state, microscopic morphology and LDC distribution of the BSA gradient solution when the concentration of the BSA solution is detected by the traditional BCA kit method, (b) is the original state, microscopic morphology and LDC distribution of the BSA gradient solution when the concentration of the BSA solution is detected using the "direct spotting-drying" strategy, (c) is the dependence between the LDC count and the gray value of the original solution and the protein concentration in the BCA kit method, and (d) is the dependence between the LDC count and the gray value of the original solution and the protein concentration under the "direct spotting-drying" strategy;

[0073] Figure 7 A schematic diagram of the data results obtained by quantitative detection of pure MnO2 provided in Example 2 of the present invention; wherein (a) is a photo of the MnO2 suspension, an ultra-depth-of-field microphotograph of the sample drop point, and an LDC count distribution diagram; (b) is a schematic diagram of the dependence of the LDC value of the sample drop point of the MnO2 suspension and the gray value of the original solution on the MnO2 concentration;

[0074] Figure 8Schematic diagram of the process and result data of quantitative detection of GSH glutathione provided in Example 3 of the present invention; wherein, (a) is a schematic diagram of MnO2 purification, (b) is a schematic diagram of the reaction of GSH glutathione and MnO2, (c) is a TEM image of a gradient suspension of pure MnO2 and a schematic diagram of LDC distribution, (d) is a schematic diagram of the dependence between LDC count and GSH concentration, and (e) is a schematic diagram of the linear fitting between LDC count and GSH concentration;

[0075] Fig. 9 A schematic diagram of the principle of quantitative detection of human serum urea provided in Example 4 of the present invention;

[0076] Fig.10 A schematic diagram of the process and result data for quantitative detection of organophosphorus pesticides provided in Example 5 of the present invention; wherein, (a) is a schematic diagram of the reaction of MnO2 with thioacetylcholine under the catalysis of acetylcholinesterase, (b) is a schematic diagram of the original state of the organophosphorus pesticide gradient solution and the corresponding LDC distribution of residual MnO2, (c) is a schematic diagram of the dependence between the concentration of organophosphorus pesticides (OPs) and the LDC count of residual MnO2, and (d) is a schematic diagram of the linear fitting results between the concentration of organophosphorus pesticides (OPs) and the LDC count of residual MnO2. DETAILED DESCRIPTION

[0077] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0078] It should be noted that, in the description of the present invention, the terms "upper", "lower", "left", "right", "inner", "outer", etc. indicating directions or positional relationships are based on the directions or positional relationships shown in the drawings, which are only for the convenience of description, and do not indicate or imply that the device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present invention. In addition, it should be noted that, in the description of the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected, or indirectly connected through an intermediate medium, or it can be the internal connection of two elements. The terms "first", "second", etc. (if any) in the present invention and the drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to the specific circumstances.

[0079] In view of the problems of complex operation and high cost of existing biochemical analysis technology, the present invention proposes a simple and efficient method for quantitative detection of biological samples based on optical discs. This detection method is based on standard optical discs and their drivers and free optical disc quality diagnosis software. Through the innovative "direct spotting-drying" (drip-dry strategy, 2D) strategy, it provides an economical and practical solution for the rapid and quantitative detection of substances in solutions. The "direct spotting-drying" strategy provided by the present invention specifically refers to directly spotting the solution to be tested on the surface of an activated optical disc. As the water evaporates, the substances in the solution will form sample spots on the surface of the optical disc. These spots will interfere with the reflection of the laser and are then recognized as error signals by the optical disc drive. There is a calibrable quantitative relationship between these error signals and the solution concentration. Combined with the optical disc quality diagnosis software, these error signals can be accurately recorded and analyzed, thereby achieving accurate quantification of the substance content in the solution.

[0080] In specific applications, any optical disc such as Blu-ray disc, CD, DVD and its corresponding optical disc drive and optical disc quality diagnosis software can be selected. Without loss of generality, the following embodiments are all described by taking Blu-ray disc as an example. Accordingly, the optical disc drive is a standard BD drive Plextor BD, and the optical disc quality diagnosis software is PlexUTILITIES. The standard BD drive is responsible for reading and writing BD disc information, while the optical disc quality diagnosis software runs on a computer and is used to analyze the quality status of the BD disc surface, especially data reading errors, such as Figure 1 shown.

[0081] Compared with CD and DVD systems, Blu-ray drives use lasers with shorter wavelengths (405nm) and objectives with larger numerical apertures, which makes the laser focus spot smaller and the storage density larger. In optical drive-based biomolecule detection, a smaller detection spot means higher detection sensitivity and resolution.

[0082] BD disc, as the core of the information storage system, has a storage capacity of up to 23.31GB. Its structure is as follows: Figure 2 As shown, the optical disc itself is a multi-layer structure, which includes, from top to bottom: hard coating 1, cover layer 2, adhesive layer 3, recording layer 4, reflective layer 5, substrate 6 and protective layer 7. The data area in the optical disc is precisely located in a circular area with an outer radius of 58mm and an inner radius of 24mm. Using sophisticated algorithms, the system can quickly convert logical locations (in GB) into physical locations (in mm) to achieve efficient data management. In addition, by using Picket code for error correction coding technology, the surface of the BD optical disc is finely divided into multiple error correction blocks of 64kb in size. These error correction blocks are arranged in an orderly manner in the form of spiral tracks, so that the sample drop points can only be presented in a radial form.

[0083] BD drives use high-precision lasers to emit lasers to accurately record and read information on BD discs. When burning data, the laser moves gradually from the inside to the outside along the spiral track on the surface of the disc, forming a series of fine pit structures on the data layer of the disc. There is a clear difference in reflectivity between these pits and the pit-free platform area on the disc, and this difference forms the basis for encoding information: when the laser irradiates the pits and the platform with the same reflectivity, it represents the binary number "0"; and when the laser scans the junction between the pits and the platform, the change in the reflection characteristics represents the binary number "1" due to the change in the direction and intensity of the reflected light.

[0084] The quality diagnostic software uses two test modes, LDC (Long Distance Code) and BIS (Burst Indicator Subcode), to monitor and quantify the number of errors within the error correction block. When the surface of a BD disc is affected by contaminants or sample droplets, it will significantly change the optical properties of the disc surface, causing the laser beam to be disturbed during the focusing process, such as Figure 2 As shown, this interference phenomenon is intuitively manifested as an LDC error count signal peak on the software interface. Each signal peak corresponds to a specific sample drop point, and the integrated area of ​​the signal peak directly reflects the LDC count caused by these drop points. The LDC count is the number of check code bytes of the LDC codeword required for error correction, reflecting the size of the error. This precise correspondence enables the present invention to achieve accurate and rapid detection of substances in solution. In practical applications, the correspondence can be calibrated by pre-preparing a standard solution.

[0085] In practical applications, the present invention will use the burning function of the Windows system to burn a 22.3GB video file into a blank BD disc through the burning function of the Windows system, so that it is full of data; then immerse it in a 0.1M NaOH solution for 1.5 hours and activate it at 55°C to change the BD surface from a hydrophobic state to a hydrophilic state. By the activation treatment, the BD surface is changed from a hydrophobic state to a hydrophilic state, and after the sample droplet is dried, it is easier to form a regular circle, which is conducive to more accurate quantitative detection. Then, as Figure 3 , Figure 4 As shown, solutions of different concentrations (preferably with a concentration gradient) are prepared as standard solutions. If an insoluble precipitate needs to be introduced, the insoluble precipitate is introduced and purified, and then the treated standard solution is dripped along the radial order of the BD surface to ensure that the solution is evenly distributed on the hydrophilic surface. If an insoluble precipitate does not need to be introduced, the standard solution is directly dripped along the radial order of the BD surface to ensure that the solution is evenly distributed on the hydrophilic surface. Subsequently, it is dried in a drying oven or at room temperature to allow the substance in the solution to be completely deposited on the BD to form a clearly identifiable sample drop point. After that, the data reading error in the BD is read using the optical disc quality diagnostic software, and the signal peak integral area of ​​each error signal is calculated to obtain the LDC count corresponding to each standard solution. Then, the concentration of the standard solution and the corresponding LDC count are fitted to obtain the corresponding curve or calculation expression, thereby realizing the calibration of the corresponding quantitative relationship.

[0086] Based on the corresponding relationship obtained by calibration, the solution to be tested or the treated solution is dropped on the corresponding diameter of the BD surface in the same way to form a sample drop point. After the water in the sample drop point is completely evaporated, the data is read, the data reading error is obtained, and the LDC count is calculated. Substituting it into the corresponding relationship, the concentration of the solution to be tested can be obtained to achieve quantitative detection.

[0087] Based on the above detection ideas provided by the present invention, Figure 5 As shown, for a single system solution that will not damage the surface of the optical disc, the "direct spotting-drying" strategy is directly applied for quantitative detection; for a complex mixed system solution or a solution that may damage the surface of the optical disc, an insoluble precipitate suspension that will not damage the surface of the optical disc is introduced, and the reaction process of the insoluble precipitate is regulated by the soluble target in the test solution, that is, the soluble target is used to react with the insoluble precipitate, or the soluble target is used to inhibit the reaction between the insoluble precipitate and other reactants to regulate the dissolution process of the insoluble precipitate, thereby achieving effective purification of the precipitate, and then the "direct spotting-drying" strategy is used to quantitatively detect the purified insoluble precipitate suspension, thereby indirectly achieving quantitative detection of the test solution.

[0088] The present invention realizes rapid, accurate and low-cost quantitative detection of substances in solution through the "direct spotting-drying" strategy. The entire detection process can be completed in only 30 minutes, and no professional detection personnel are required, which further reduces the cost and time cost of biomacromolecule detection. In addition, by regulating the dissolution process of insoluble precipitates through soluble targets, indirect quantitative detection of substances in complex mixed system solutions can also be achieved, broadening its scope of application.

[0089] The following is further explained with reference to specific embodiments.

[0090] Embodiment 1:

[0091] The invention discloses a biological sample quantitative detection method based on an optical disk, which is used to realize the quantitative detection of a bovine serum albumin (BSA) solution.

[0092] This example also uses the traditional BCA kit method to perform quantitative detection on the same bovine serum albumin solution, and its detection process and detection results serve as a comparative example of this example.

[0093] Figure 6 (a) shows the original solution state of the protein gradient solution with the addition of BCA reagent when using the traditional BCA kit method. The microscopic morphology of the sample drop point is captured through an ultra-depth of field micrograph, and the LDC distribution is displayed. Figure 6 (c) in the figure intuitively reflects the dependence between protein LDC counts and grayscale values ​​of the original solution and protein concentration in the BCA method.

[0094] Figure 6 The photograph of the reaction solution shown in (a) clearly shows the significant color gradient changes formed by different concentrations of BSA. After grayscale value analysis using ImageJ software, the results showed that there was an acceptable linear relationship between grayscale value and BSA concentration. However, after applying the drip-drying strategy, it was found that the LDC distribution remained roughly at the same level, as shown in Figure 2. Figure 6 As shown in (c), the ultra-depth micrographs also show that although the pattern changes slightly, there is almost no significant difference in the coverage of each sample point. This observation is likely due to an important fact in the BCA colorimetric method: the concentration of the detection reagent is much higher than the concentration of BSA. Therefore, during the evaporation and drying process, all solutes (including detection reagents and BSA) will remain on the BD surface. This shows that the drip-drying strategy does not rely on color changes in traditional colorimetric methods, but more on the total concentration of all substances in the solution.

[0095] Subsequently, pure BSA gradient solutions were prepared in this example, and the above-mentioned "direct spotting-drying" strategy was used to calculate the LDC count. Although the concentration differences of these solutions were difficult to distinguish visually, the LDC distribution at the sample drop point showed an obvious gradient trend, as shown in Figure 2. Figure 6 As shown in (b) in Figure 2, through calculation, it was found that there was a positive correlation between LDC count and BSA concentration, as shown in Figure 6 Although a saturation trend was observed at higher BSA concentrations, this still demonstrates the potential of the drip-drying strategy for quantitative detection of substances in a single system within a specific concentration range. It should be noted that this specific concentration range is sufficient to cover the concentration of BSA solution that is actually required to be measured.

[0096] by Figure 6 The curve obtained by fitting the BAS concentration and the corresponding LDC count shown in (d) is the standard curve. Based on the standard curve, the quantitative detection method of bovine serum albumin using the "direct spotting-drying" strategy provided by the present invention in this embodiment specifically includes:

[0097] After the optical disc is activated, the bovine serum protein solution to be tested is dripped on the surface of the optical disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the bovine serum protein solution to be tested is the same as the diameter of the standard solution when calibrating the standard curve;

[0098] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0099] The concentration corresponding to the calculated LDC counts is determined according to a pre-calibrated standard curve to obtain the concentration of the bovine serum albumin solution to be tested.

[0100] It should be noted that if the relevant standard curve has been calibrated in advance, there is no need to calibrate it, and the detection can be directly performed based on the calibrated standard curve.

[0101] By comparison, it was found that when using the traditional BCA kit for protein quantification, the protein needs to react with the colorimetric reagent at room temperature for 1-2 hours before the absorbance can be measured or the colorimetric analysis can be performed. However, this embodiment completely eliminates this cumbersome reaction step and can directly quantify the pure protein in about 20 minutes. This result fully confirms the great potential of the CD-based biological sample quantitative detection method assisted by the "drip-dry strategy, 2D" strategy in rapid POCT (point-of-care testing).

[0102] Embodiment 2:

[0103] A method for quantitative detection of biological samples based on optical disks is used to achieve quantitative detection of MnO2 solution. This embodiment also adopts the above-mentioned "direct spotting-drying" strategy to directly perform quantitative detection on the MnO2 solution to be tested.

[0104] In order to calibrate the standard curve for recording the corresponding relationship between the concentration of MnO2 solution and the LDC count, this embodiment uses deionized water and MnO2 nanoparticles (particle size between 100nm and 150nm) to prepare 8 suspensions with different concentration gradients in 8 EP tubes (microcentrifuge tubes), and the concentrations are: 0, 0.1mg / mL, 0.2mg / mL, 0.3mg / mL, 0.4mg / mL, 0.6mg / mL, 0.8mg / mL, 1.0mg / mL). Then, these EP tubes are placed in an ultrasonic cleaner for cleaning for 15 minutes to ensure that the MnO2 nanoparticles are evenly distributed in the suspension. Subsequently, the obtained MnO2 gradient suspension is uniformly dripped on the surface of the Blu-ray disc after activation treatment along the specified radius in sequence to form corresponding sample drop points. After the water in each sample drop point is completely evaporated, the BD error is read to obtain the data reading error corresponding to the MnO2 gradient suspension, and the LDC of each signal peak is calculated and the gray value analysis is performed. The results of these analyses are presented in the form of photographs of the MnO2 gradient suspension, ultra-depth-of-field micrographs of the sample drop point, and LDC distribution diagrams, such as Figure 7 As shown in (a) in the figure. The LDC value of the sample drop point and the gray value of the original solution were further plotted and analyzed, thereby obtaining the dependence of the LDC value and gray value on the solution concentration, as shown in Figure 7 As shown in (b) in .

[0105] Generally speaking, the results of grayscale value analysis are more accurate, but the analysis is time-consuming and costly. On the contrary, the analysis based on LDC count is fast and cost-effective. Figure 7 Taking the fitting curve of the gray value and solution concentration shown in (b) as a reference, it can be found that the fitting curve of LDC count and solution concentration can more accurately reflect the actual solution concentration. In other words, this embodiment can effectively improve the detection efficiency and reduce the detection cost while ensuring the detection accuracy.

[0106] based on Figure 7 The standard curve shown in (b) is used to record the corresponding relationship between the concentration of MnO2 solution and the LDC count. The quantitative detection method of MnO2 solution based on the "direct spotting-drying" strategy in this embodiment specifically includes:

[0107] After the optical disc is activated, the MnO2 solution to be tested is dripped on the surface of the optical disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the drop of the MnO2 solution to be tested is the same as the diameter of the drop of the standard solution when calibrating the standard curve;

[0108] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0109] The concentration corresponding to the calculated LDC counts is determined according to a pre-calibrated standard curve to obtain the concentration of the MnO2 solution to be tested.

[0110] It should also be noted that if the relevant standard curve has been calibrated in advance, there is no need to calibrate it, and the detection can be directly performed based on the calibrated standard curve.

[0111] Embodiment 3:

[0112] The invention discloses a biological sample quantitative detection method based on an optical disk, which is used to realize the quantitative detection of a GSH glutathione solution.

[0113] GSH is a non-protein thiol formed by the combination of glutamate, cysteine ​​and glycine. It acts as an antioxidant in the human body to maintain cellular redox homeostasis. GSH exists mainly in a reduced state in the human body and can be oxidized by MnO2 to generate oxidized glutathione GSSG, such as Figure 8 As shown in (b) in .

[0114] In order to detect the concentration of GSH solution, this example firstly reacted a series of GSH solutions of graded concentrations with an excess MnO2 (2 mg / mL) solution of equal concentration. After the mixed liquid was completely reacted, it was centrifuged so that the remaining MnO2 was deposited at the bottom of the centrifuge tube. Then, the supernatant was removed, that is, the GSSG and Mn produced by the reaction were removed. 2+ , then add deionized water to resuspend MnO2, thereby obtaining a set of pure MnO2 gradient suspensions, such as Figure 8 As shown in (a) in .

[0115] The obtained pure MnO2 gradient suspension is uniformly dripped on the surface of the activated Blu-ray disc along the specified radius to form a sample drop point. Then, the data of the Blu-ray disc is read to obtain the corresponding data reading error, such as Figure 8 As shown in (c) in the figure; since GSH can react directly with MnO2, the higher the GSH content, the higher the amount of MnO2 consumed, which leads to a corresponding decrease in the turbidity of the MnO2 suspension. Figure 8 The result shown in (c) is consistent with this. At the same time, this example performed transmission electron microscopy (TEM) imaging on the MnO2 suspension before and after the reaction. The TEM image shows that the density of MnO2 particles decreases after the reaction, which further proves that GSH has the property of dissolving MnO2.

[0116] Next, we used the "drip-dry strategy" (2D strategy) to perform BD error reading on the obtained MnO2 suspension and calculate the LDC counts of each signal peak. The results showed that the LDC counts of residual MnO2 were negatively correlated with the GSH concentration, such as Figure 8 As shown in (d) in the figure. Further analysis showed that the LDC count and GSH concentration showed a good linear relationship in the range of 2-16 mM. The specific formula is LDC count = (-1.951 × [GSH] + 5.850) × 10 3 , linear fit R 2 =0.991, such as Figure 8 As shown in (e) in .

[0117] It was calculated that the detection limit (LOD) of GSH in this method was about 0.447 mM, which preliminarily met the requirements for GSH detection in normal human cells (usually ranging between 1-10 mM).

[0118] Based on the above relationship between the concentration of GSH solution and LDC count, the quantitative detection method of GSH solution based on the "direct spotting-drying" strategy in this embodiment specifically includes:

[0119] The GSH solution to be tested was mixed with an excess MnO2 solution, and after GSH and MnO2 were fully reacted, the mixed solution was centrifuged; the concentration of the MnO2 solution was the same as that of the MnO2 solution selected during calibration, both of which were 2 mg / mL;

[0120] After removing the supernatant in the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new MnO2 suspension, and the MnO2 suspension is dripped on the surface of the activated Blu-ray disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the dripping MnO2 suspension is the same as the diameter of the dripping standard solution when calibrating the relationship;

[0121] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0122] Substituting the LDC count into the above relationship between the GSH solution concentration and the LDC count, the solubility of the GSH solution to be tested is obtained.

[0123] This example provides a new idea for GSH detection. It is expected that the experimental conditions can be further optimized and the parameters can be adjusted in the future to further reduce the detection limit.

[0124] It is easy to understand that when the selected MnO2 solution changes, the relevant fitting relationship will change, and it can be recalibrated according to the corresponding calibration method. It should also be noted that if the relevant relationship has been calibrated, there is no need to calibrate it, and the detection can be directly based on the calibrated relationship.

[0125] Embodiment 4:

[0126] A biological sample quantitative detection method based on an optical disk is used to achieve quantitative detection of serum uric acid solution.

[0127] Serum uric acid is the final product of purine catabolism and is mainly excreted from the body through urine by the kidneys. Under normal physiological conditions, the concentration of serum uric acid in men is usually in the range of 149-416 μmol / L (or 150-420 μmol / L), while that in women is 89-357 μmol / L (or 90-357 μmol / L).

[0128] Uric acid is reducing. When it comes into contact with manganese dioxide (MnO2), uric acid will be oxidized, while MnO2 itself will be reduced to produce allantoin and Mn 2+ ,like Fig. 9 As shown. The detection principle of this embodiment is the same as that of the above-mentioned embodiment 3. In order to use the above-mentioned "direct spotting-drying" strategy to detect the content of serum uric acid, this embodiment prepared a group of serum uric acid solutions with different concentration gradients, and made them react separately with excess MnO2 of equal concentration. After the reaction is complete, the mixed liquid is centrifuged so that the remaining MnO2 in the reaction is deposited at the bottom of the centrifuge tube. Subsequently, the supernatant is removed, that is, the allantoin and Mn produced during the reaction are removed. 2+ , and then deionized water is added to resuspend the MnO2 suspension to obtain a set of pure MnO2 gradient suspensions. In this process, since serum uric acid directly reacts with MnO2, the higher the serum uric acid content, the higher the amount of MnO2 consumed, resulting in a corresponding decrease in the turbidity of the MnO2 suspension.

[0129] Next, the obtained MnO2 suspension is subjected to BD error reading, and the LDC count of each signal peak is calculated. These data are then converted into a concentration curve for analyzing and evaluating the content of serum uric acid. Similar to the above-mentioned Example 3, this embodiment can calibrate a relationship for the corresponding relationship between serum uric acid concentration and LDC count. Based on the calibrated relationship, the quantitative detection method of serum uric acid solution based on the "direct spotting-drying" strategy in this embodiment specifically includes:

[0130] The serum uric acid solution to be tested is mixed with an excess MnO2 solution, and after the uric acid and MnO2 are fully reacted, the mixed solution is centrifuged;

[0131] After removing the supernatant in the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new MnO2 suspension, and the MnO2 suspension is dripped on the surface of the activated Blu-ray disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the dripping MnO2 suspension is the same as the diameter of the dripping standard solution when calibrating the relationship;

[0132] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0133] Substituting the LDC count into the above relationship between the serum uric acid solution concentration and the LDC count, the solubility of the GSH solution to be tested is obtained.

[0134] Embodiment 5:

[0135] A biological sample quantitative detection method based on an optical disk is used to achieve quantitative detection of an organophosphorus pesticide solution.

[0136] Existing counting methods often use traditional capillary electrophoresis (CE), high performance liquid chromatography (HPLC) and gas chromatography (GC) to achieve quantitative detection of organophosphorus pesticides (OPs). These detection methods have high detection sensitivity, but rely on well-trained professionals and expensive laboratory equipment and are not suitable for on-site analysis.

[0137] At present, a popular rapid on-site detection method is to combine colorimetry and enzyme inhibition. In this method, the reaction process of acetylcholinesterase (AChE) catalyzing the hydrolysis of thioacetylcholine (ATCh) to generate thiocholine (TCh) is used, and the inhibition of the catalytic function of acetylcholinesterase (AChE) by organophosphorus pesticides, thereby affecting the subsequent color development process. On this basis, this embodiment uses MnO2 as a common oxidant to react with thioacetylcholine (ATCh) to form water-soluble Mn 2+ ,like Fig.10 As shown in (a) in .

[0138] Since organophosphorus pesticides can inhibit the catalytic function of acetylcholinesterase (AChE), the presence of organophosphorus pesticides can inhibit the dissolution of MnO2. Therefore, this embodiment can indirectly evaluate the concentration of organophosphorus pesticides by detecting the residual amount of MnO2. In order to detect the concentration of the organophosphorus pesticide solution using the "direct spotting-drying" strategy, this embodiment first mixes a series of graded concentrations of organophosphorus pesticides with equal concentrations and excess MnO2 suspensions, thioacetylcholine solutions, and acetylcholinesterase solutions, and allows each mixed solution to react separately. After the mixed liquid is completely reacted, it is centrifuged so that the remaining MnO2 in the reaction is deposited at the bottom of the centrifuge tube. Next, the supernatant is removed, and deionized water is added to resuspend the MnO2 suspension to obtain a group of pure MnO2 gradient suspensions. Since the presence of organophosphorus pesticides can inhibit the dissolution of MnO2, the higher the content of organophosphorus pesticides, the higher the residual amount of MnO2, and the turbidity of the MnO2 suspension also increases accordingly, such as Fig.10 As shown in (b) in .

[0139] The obtained pure MnO2 gradient suspension was evenly dripped on the surface of the activated Blu-ray disc along the specified radius to form a sample drop point. The data of the Blu-ray disc was read to obtain the corresponding data reading error, and the LDC count of each signal peak was calculated. It was found that the LDC count of residual MnO2 was positively correlated with the concentration of organophosphorus pesticides (OPs), such as Fig.10 As shown in (c) of Figure 2, further analysis showed that the LDC count and OPs concentration were in the range of 1.289×10 -7 -1.289×10 -4 There is an acceptable linear relationship within the range of M. The specific formula is LDC count = (0.608 × lg [OPs] + 4.386) × 10 3 , linear fit R 2 =0.978, such as Fig.10 In addition, the detection limit of this method for organophosphorus pesticides is about 2.122×10 - 7 M, to meet the actual detection needs.

[0140] It is easy to understand that when the selected MnO2 solution changes, the relevant fitting relationship will change, and it can be recalibrated according to the corresponding calibration method. Therefore, the dynamic range and detection limit can be further optimized by adjusting the reaction time and MnO2 concentration.

[0141] Based on the above relationship between the concentration of the organophosphorus pesticide solution and the LDC count, the quantitative detection method of the organophosphorus pesticide solution based on the "direct spotting-drying" strategy in this embodiment specifically includes:

[0142] The organophosphorus pesticide solution to be tested is mixed with an excess of MnO2 suspension, thioacetylcholine solution, and acetylcholinesterase solution, and after the mixed solution is fully reacted, the mixed solution is centrifuged;

[0143] After removing the supernatant in the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new MnO2 suspension, and the MnO2 suspension is dripped on the surface of the activated Blu-ray disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the dripping MnO2 suspension is the same as the diameter of the dripping standard solution when calibrating the relationship;

[0144] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0145] Substitute the LDC count into the above relationship between the concentration of the organophosphorus pesticide solution and the LDC count to obtain the solubility of the organophosphorus pesticide solution to be tested.

[0146] It should also be noted that if the relevant relationship has been calibrated, there is no need to calibrate it, and the detection can be directly performed based on the calibrated relationship.

[0147] Embodiment 6:

[0148] A biological sample quantitative detection method based on an optical disk is used to achieve quantitative detection of bacterial concentration.

[0149] Bacteria concentration detection plays an indispensable role in many key fields such as medicine, food, and environment, and is of great significance for early detection of diseases, ensuring food safety, and evaluating environmental hygiene. However, traditional bacteria detection methods are difficult to meet the needs of rapid on-site monitoring due to limitations such as complex operations and high costs.

[0150] The detection principle of this embodiment is similar to that of the above-mentioned embodiments 1 and 2. In order to detect the bacterial concentration based on the above-mentioned "direct spotting-drying" strategy, this embodiment first prepares a series of standard bacterial solutions with different concentrations by precise dilution with deionized water. Then, the Blu-ray disc is subjected to a specific activation treatment, which causes the surface of the disc to change from a hydrophobic state to a hydrophilic state. At the same time, after the activation treatment, the ability of the disc surface to adsorb bacteria will be enhanced. Then, the standard bacterial solution to be tested is evenly dripped on the surface of the activated Blu-ray disc along a specified radius to form a sample drop point. As the water gradually evaporates, the bacteria in each sample drop point will be deposited on the surface of the disc to form a clearly identifiable spot point. After that, the data of the Blu-ray disc is read to obtain the read data error corresponding to each sample drop point. And further calculate the LDC count of each signal peak in the error signal. These counts reflect the density and distribution of bacterial deposition on the surface of the disc. Based on the calculated LDC count, the concentration of each standard bacterial solution and the corresponding LDC count are fitted, and a standard curve for recording the corresponding relationship between concentration and LDC count can be calibrated.

[0151] Based on the calibrated standard curve, this embodiment uses the "direct spotting-drying" strategy provided by the present invention to quantitatively detect the bacterial solution, specifically including:

[0152] After the optical disc is activated, a bacterial solution is dripped on the surface of the optical disc at a specified radius to form a sample drop point; it is easy to understand that the diameter of the bacterial solution drop is the same as the diameter of the standard solution drop when calibrating the standard curve;

[0153] After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count;

[0154] The concentration corresponding to the calculated LDC counts was determined according to a pre-calibrated standard curve to obtain the concentration of the bacterial solution.

[0155] The quantitative detection method of bacterial solution provided in this embodiment is not only fast but also highly accurate, and is applicable to various bacterial detection scenarios, providing an innovative method for rapid in vitro detection of bacteria.

[0156] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A biological sample quantitative detection method based on an optical disc, for detecting the concentration of a solution to be tested; the solution to be tested is a soluble target solution or an insoluble precipitate suspension, and the soluble target of the soluble target solution and the insoluble precipitate in the insoluble precipitate suspension will not damage the surface of the optical disc; The optical disc is filled with data; it is characterized in that: The quantitative detection method comprises: After the optical disc is activated, the solution to be tested is dripped on a specified radius on the surface of the optical disc to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state; After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error therein, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count; Determine the concentration corresponding to the calculated LDC count according to a pre-calibrated standard curve to obtain the concentration of the test solution; Wherein, when the test solution is a soluble target solution, the standard curve is obtained by fitting the LDC counts corresponding to soluble target solutions of different concentrations; when the test solution is an insoluble precipitate suspension, the standard curve is obtained by fitting the LDC counts corresponding to insoluble precipitate suspensions of different concentrations.

2. The method for quantitative detection of biological samples based on an optical disk according to claim 1, characterized in that: The calibration method of the standard curve includes: Prepare test solutions with different concentrations as standard solutions; After the optical disc is activated, the standard solution is uniformly dripped on the surface of the optical disc along the specified radius to obtain a sample drop point corresponding to each standard solution; After the water in all the sample drop points is completely evaporated, the data in the optical disc is read to obtain the data reading error corresponding to each sample drop point, and the integrated area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution; Fitting was performed based on the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the corresponding relationship between the concentration and the LDC count.

3. A method for quantitative detection of biological samples based on an optical disk, for detecting the concentration of a test solution, wherein the test solution is a soluble target solution; The optical disc is filled with data; it is characterized in that: The quantitative detection method comprises: Step S1: mixing a test solution with an excess of an insoluble precipitate suspension, so that the soluble target in the test solution and the insoluble precipitate in the insoluble precipitate suspension fully react; the soluble target can react with the insoluble precipitate to produce a soluble product after the reaction; the insoluble precipitate will not damage the surface of the optical disc; Step S2: centrifuging the mixed solution; Step S3: after removing the supernatant from the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new insoluble precipitate suspension; Step S4: after the optical disc is activated, the insoluble precipitate suspension obtained in step S3 is dripped on the surface of the optical disc at a specified radius to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state; Step S5: after the water in the sample droplet point is completely evaporated, the data in the optical disc is read to obtain the data reading error therein, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count; Step S6: Substitute the LDC count obtained in step S5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

4. The method for quantitative detection of biological samples based on an optical disk according to claim 3, characterized in that: The calibration method of the relationship between LDC count and concentration of the solution to be tested includes: Prepare test solutions with different concentrations as standard solutions; Each standard solution is mixed with an insoluble precipitate suspension of equal concentration and excess, so that the soluble target in each standard solution and the insoluble precipitate in the insoluble precipitate suspension can fully react; Each mixed solution is centrifuged separately; After removing the supernatant from the mixed solution after centrifugation, deionized water was added to the remaining solution to obtain a new insoluble precipitate suspension corresponding to each standard solution; After the optical disc is activated, the insoluble precipitate suspension corresponding to each standard solution is uniformly dropped on the surface of the optical disc along the specified radius to obtain a sample dropping point corresponding to each standard solution; After the water in all the sample drop points is completely evaporated, the data in the optical disc is read to obtain the data reading error corresponding to each sample drop point, and the integrated area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution; The concentration of each standard solution and the corresponding LDC count were fitted to obtain the relationship between the LDC count and the concentration of the solution to be tested.

5. A method for quantitative detection of biological samples based on an optical disk, for detecting the concentration of a solution to be tested, wherein the solution to be tested is a soluble target solution; The optical disc is filled with data; it is characterized in that: The quantitative detection method comprises: Step T1: mixing a test solution, an insoluble precipitate suspension and a third reactant solution, so that the insoluble precipitate in the insoluble precipitate suspension fully reacts with the third reactant in the third reactant solution; the insoluble precipitate can react with the third reactant to produce a soluble product after the reaction, and the insoluble precipitate suspension is in excess relative to the third reactant solution; the test solution will inhibit the reaction of the third reactant with the insoluble precipitate, and the higher the concentration of the test solution, the stronger the inhibitory effect; the insoluble precipitate will not damage the surface of the optical disc; Step T2: centrifuging the mixed solution; Step T3: after removing the supernatant from the mixed solution after centrifugation, deionized water is added to the remaining solution to obtain a new insoluble precipitate suspension; Step T4: after the optical disc is activated, the insoluble precipitate suspension obtained in step T3 is dripped on a specified radius on the surface of the optical disc to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state; Step T5: after the water in the sample dripping clock is completely evaporated, the data in the optical disc is read to obtain the data reading error therein, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count; Step T6: Substitute the LDC count obtained in step T5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

6. The method for quantitative detection of biological samples based on an optical disk according to claim 5, characterized in that: The calibration method of the relationship between LDC count and concentration of the solution to be tested includes: Prepare test solutions with different concentrations as standard solutions; For each standard solution, mix it with the insoluble precipitate suspension and the third reactant solution, so that the insoluble precipitate and the third reactant fully react; the concentration of the insoluble precipitate suspension mixed with each standard solution is equal, and the concentration of the third reactant solution mixed with each standard solution is equal; Each mixed solution is centrifuged separately; After removing the supernatant from the mixed solution after centrifugation, deionized water was added to the remaining solution to obtain a new insoluble precipitate suspension corresponding to each standard solution; After the optical disc is activated, the insoluble precipitate suspension corresponding to each standard solution is uniformly dropped on the surface of the optical disc along the specified radius to obtain a sample dropping point corresponding to each standard solution; After the water in all the sample drop points is completely evaporated, the data in the optical disc is read to obtain the data reading error corresponding to each sample drop point, and the integrated area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution; The concentration of each standard solution and the corresponding LDC count were fitted to obtain the relationship between the LDC count and the concentration of the solution to be tested.

7. A method for quantitative detection of biological samples based on an optical disc, for detecting the concentration of a bacterial solution; the optical disc is filled with data; characterized in that: The quantitative detection method comprises: After the optical disc is activated, the bacterial solution is dripped on a specified radius on the surface of the optical disc to form a sample drop point; the activation treatment causes the surface of the optical disc to change from a hydrophobic state to a hydrophilic state; After the water in the sample drop point is completely evaporated, the data in the optical disc is read to obtain the data reading error therein, and the integral area of ​​the signal peak of the data reading error is calculated to obtain the corresponding LDC count; Determine the concentration corresponding to the calculated LDC count according to a pre-calibrated standard curve to obtain the concentration of the bacterial solution; The standard curve is obtained by fitting the LDC counts corresponding to bacterial solutions of different concentrations.

8. The method for quantitative detection of biological samples based on an optical disk according to claim 7, characterized in that: The calibration method of the standard curve includes: Prepare bacterial solutions with different concentrations as standard solutions; After the optical disc is activated, the standard solution is uniformly dripped on the surface of the optical disc along the specified radius to obtain a sample drop point corresponding to each standard solution; After the water in all the sample drop points is completely evaporated, the data in the optical disc is read to obtain the data reading error corresponding to each sample drop point, and the integrated area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution; Fitting was performed based on the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the corresponding relationship between the concentration and the LDC count.

9. The method for quantitative detection of biological samples based on an optical disk according to any one of claims 1 to 8, characterized in that: The activation treatment comprises: The optical disc is immersed in a NaOH solution for a preset time to generate reactive carboxyl groups on its surface, thereby changing from a hydrophobic state to a hydrophilic state.

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